RNF2 is the target for phosphorylation by the p38 MAPK and ERK signaling pathways.
RNF2 is the target for phosphorylation by the p38 MAPK and ERK signaling pathways.
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DOI：
10.1002/pmic.200800847
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发表时间：
2009-05
期刊：
影响因子：
3.4
通讯作者：
Rao, U. Subrahmanyeswara
中科院分区：
文献类型：
--
作者：
Rao, Prema S.;Satelli, Arun;Zhang, Sheng;Srivastava, Sanjay K.;Srivenugopal, Kalkunte S.;Rao, U. Subrahmanyeswara
RNF2 是多梳族 (PcG) 蛋白的成员，参与染色质重塑。然而，调节 RNF2 功能的机制尚不清楚。为了确定此类机制，RNF2 在 HEK-293 细胞中表达并通过二维电泳进行分析。 RNF2 被分解为至少 7 个蛋白点，从预期的 pI 6.38 向较低的 pI 迁移，表明 RNF2 经历了翻译后修饰。蛋白质印迹表明，这些 RNF2 斑点中的大多数含有磷酸丝氨酸，在用磷酸酶处理后完全去磷酸化。 SB203580 是 p38 MAPK 的一种特异性抑制剂，可抑制 RNF2 在一个位点的磷酸化。另一方面，MEK1/2 抑制剂 PD98059 抑制 RNF2 中的大部分磷酸化事件。质谱分析发现，Sf9 昆虫细胞中表达的 RNF2 经历了 1Met 的共翻译切除、2Ser 的 N 乙酰化和 41Ser 的磷酸化。有趣的是，41Ser 是预测的 p38/MAPK 磷酸化位点，与 SB203580 诱导的磷酸化丧失一致。进一步分析表明，RNF2 磷酸化差异调节转录因子和组蛋白 2B 乙酰化的表达。这些结果为RNF2磷酸化提供了第一个证据，并表明包括p38 MAPK和ERK1/2在内的丝裂原激活蛋白激酶通过RNF2磷酸化调节生长、应激反应、分化和其他细胞过程。
RNF2, a member of polycomb group (PcG) proteins, is involved in chromatin remodeling. However, mechanisms that regulate RNF2 function are unknown. To identify such mechanisms, RNF2 was expressed in HEK‐293 cells and analyzed by 2‐D electrophoresis. RNF2 was resolved into at least seven protein spots, migrating toward the lower pIfrom its expected pIof 6.38, suggesting that RNF2 undergoes post‐translational modifications. Western blotting indicated that majority of these RNF2 spots contained phosphoserine(s), which were completely dephosphorylated upon treatment with a phosphatase. SB203580, a specific inhibitor of p38 MAPK, inhibited RNF2 phosphorylation at one site. On the other hand, PD98059, an inhibitor of MEK1/2, inhibited majority of the phosphorylation events in RNF2. Mass spectrometry analysis identified that RNF2 expressed in Sf9 insect cells undergoes co‐translational excision of1Met coupled to N‐acetylation of2Ser, and phosphorylation of41Ser. Interestingly,41Ser is a predicted p38/MAPK phosphorylation site, consistent with the loss of phosphorylation induced by SB203580. Further analysis indicated that RNF2 phosphorylation differentially modulates the expression of transcription factors and histone 2B acetylation. These results provide first evidence for phosphorylation of RNF2, and suggest that the mitogen activated protein kinases including p38 MAPK and ERK1/2 regulate growth, stress response, differentiation and other cellular processes, through phosphorylation of RNF2.
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